Convenient assay for interferons

Rubinstein, Sara; Familletti, Philip C.; Pestka, Sidney

doi:10.1128/jvi.37.2.755-758.1981

Cited by 453 publications

(109 citation statements)

References 10 publications

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“…Virus titers were calculated by determining the dilution giving 50% of wells containing cells that displayed cytopathic effect. Antiviral activity was determined with a standard cytopathic effect assay (Rubinstein et al, 1981) with some modifications (Boue et al, 2000;Bracklein et al, 2006;Rodriguez et al, 1998b;Shao et al, 2015a). Briefly, the monolayers cells were seeded in 96 well plates, then inoculated with four-fold serial diluted BoIFNs.…”

Section: Antiviral Activity and Antibody Blocking Assay In Vitromentioning

confidence: 99%

Molecular characterization and biological activity of bovine interferon-omega3

Guo

Bao

et al. 2017

Research in Veterinary Science

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Bovine interferon-omega3 (BoIFN-ω3) gene was amplified from bovine liver genomic DNA, which encodes a 195-amino acid protein containing a 23-amino acid signal peptide. Analysis of the molecular characteristics revealed that BoIFN-ω3 evolving from IFN-ω, contained four cysteine residues and five alpha helices, showing that BoIFN-ω3 presented the typical molecular characteristics of type I interferon. BoIFN-ω3 exhibited antiviral and antiproliferative activities, which exerted a protective effect against VSV in several mammalian cell lines, as well as against BEV, IBRV, and BVDV in MDBK cell. Moreover, BoIFN-ω3 was shown to be highly sensitive to trypsin, but remaining stable despite changes in pH and temperature. Additionally, BoIFN-ω3 induced the transcription of Mx1, ISG15, and ISG56 genes, as well as the expression of Mx1 protein in a time-dependent manner. These findings will be useful to further study BoIFN-ω in host's defence against infectious diseases, particularly viral infections. Furthermore, results will facilitate further research on the bovine interferon family.

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Section: Antiviral Activity and Antibody Blocking Assay In Vitromentioning

confidence: 99%

Molecular characterization and biological activity of bovine interferon-omega3

Guo

Bao

et al. 2017

Research in Veterinary Science

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“…The IFN-a bioassay was modified from that reported by Rubenstein et al (1981). Before the measurement, the culture supernatants collected at various incubation intervals were thawed and irradiated with UV light at a distance of 15 cm for 20 min by using a GL-15 UV lamp (15 W, Toshiba, Tokyo, Japan) to inactivate the remaining PCV2 and PRRSV.…”

Section: Interferon-alpha Bioassaymentioning

confidence: 99%

Reduction of porcine reproductive and respiratory syndrome virus (PRRSV) infection in swine alveolar macrophages by porcine circovirus 2 (PCV2)-induced interferon-alpha

Chang

Jeng

Liu

et al. 2005

Veterinary Microbiology

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Two common viral pathogens of swine, namely, porcine circovirus type 2 (PCV2) and porcine reproductive and respiratory syndrome virus (PRRSV), were investigated in regard to their effects on monolayer cultures of swine alveolar macrophages (AMs). The purpose was to identify selected cellular changes and responses potentially associated with the clinical reactions of pigs infected with either or both of these viruses. Measurements included the (1) absolute and relative numbers of infected, viable, and apoptotic cells; (2) distribution of viral antigens; (3) levels of interferon-alpha (IFN-alpha) and tumor necrosis factor-alpha (TNF-alpha) produced and their association with the extent of virus-induced cytopathology. Four groups of AMs were studied, including mock-infected, PCV2 alone-infected (PCV2-A), PRRSV alone-infected (PRRSV-A), and PCV2 and PRRSV dually infected (PCV2/PRRSV) groups. The AMs of PCV2-A group had high antigen-containing rate without cell death. There was a marked increase in cell death and apoptosis in PRRSV-A group. However, a lower PRRSV-induced infectious rate, cell death, and apoptosis were seen in PCV2/PRRSV group. High levels of IFN-alpha production were detected in PCV2-infected groups, but not in mock-infected and PRRSV-A groups. The PRRSV-induced cytopathic effect (CPE) on MARC-145 cells or swine AMs was markedly reduced by pre-incubation of the cells with UV-treated or non-UV-treated supernatants of PCV2-infected AMs. In addition, the reduction in CPE was abolished when the supernatants of PCV2-infected AMs were pre-treated with a mouse anti-recombinant porcine IFN-alpha antibody. The results suggest that swine AMs were an important reservoir of PCV2; PCV2 infection reduced PRRSV infection and PRRSV-associated CPE in PCV2/PRRSV AMs; the reduction of PRRSV infection in AMs was mediated by IFN-alpha generated by PCV2 infection. The reduced PRRSV-associated CPE in AMs and increased pro-inflammatory cytokine production may lead to a more severe pneumonic lesion in those dually infected pigs.

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“…To verify that an active cytokine was encoded by the amplified cDNA, Chinese hamster ovary (CHO) cells were transfected with pINA and single cell clones resistant to 1.0 mg geneticin per ml growth medium were prepared. Supernatant medium from the clones were tested for the ability to inhibit the replication of an interferoninducer negative strain of vesicular stomatitis virus (Sekellick and Marcus, 1979) in Madin Derby bovine kidney (MDBK) cells as previously described (Rubinstein et al, 1981). Clones producing from 0 to greater than 200,000 units (1 unit inhibits 50% of VSV replication in MDBK cell monolayers) of IFN-a were detected.…”

Section: Mammalian Expression Vectors Containing Porcine Ifn-a or Il-mentioning

confidence: 99%

Cytokines and synthetic double-stranded RNA augment the T helper 1 immune response of swine to porcine reproductive and respiratory syndrome virus

Meier

Husmann

Schnitzlein

et al. 2004

Veterinary Immunology and Immunopathology

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Immunization of pigs with a modified live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine initially elicits a weak interferon (IFN)-gamma response. To improve the immune response, an adjuvant consisting of plasmid encoding either porcine interleukin (IL)-12 or IFN-alpha was co-administered during vaccination. In the presence of either adjuvant, at least a three-fold increase in the primary virus-specific IFN-gamma response was observed. While this enhancement was only transient (1 week) when the IL-12 expressing plasmid was used, the effect was not only still apparent at 6 weeks after vaccination in the presence of the IFN-alpha expressing plasmid but even after challenge with a virulent genetically divergent PRRSV. In contrast, no effect of either adjuvant on the production of anti-virus antibodies was noticed throughout the study. Despite the apparent augmentation of a T helper (Th) 1 type response by the inclusion of IFN-alpha or IL-12 during vaccination, this modulation did not necessarily correlate with a reduction in viremia. Since a similar increase in the degree of the IFN-gamma response to the PRRSV vaccine could be achieved by substituting polyinosinic-polycytidylic acid in lieu of either cytokine, exposure to PRRSV in the presence of a variety of Th 1 polarizing molecules can positively influence the development of the cell-mediated immune response of swine to this pathogen. Conceivably, such intervention could be applied to improve the formulation of anti-PRRSV vaccines.

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Convenient assay for interferons

Cited by 453 publications

References 10 publications

Molecular characterization and biological activity of bovine interferon-omega3

Molecular characterization and biological activity of bovine interferon-omega3

Reduction of porcine reproductive and respiratory syndrome virus (PRRSV) infection in swine alveolar macrophages by porcine circovirus 2 (PCV2)-induced interferon-alpha

Cytokines and synthetic double-stranded RNA augment the T helper 1 immune response of swine to porcine reproductive and respiratory syndrome virus

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